Anti mll2

Manufactured by Fortis Life Sciences

Anti-MLL2 is a laboratory reagent used for the detection and analysis of the MLL2 (Lysine-specific methyltransferase 2D) protein in biological samples. MLL2 is an enzyme involved in the epigenetic regulation of gene expression. Anti-MLL2 can be used in various research applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and localization of the MLL2 protein.

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3 protocols using anti mll2

Comprehensive Histone Modification Profiling

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The following antibodies were used in this study: anti-H3K4me1 (generated in-house), anti-H3K4me2 (generated in-house), anti-H3K4me3 (generated in-house), anti-H3K27ac (Cell Signaling, 8173), anti-H3 (generated in-house), anti-SMC1A (Bethyl Laboratories, A300-055A), anti-SET1A (generated in-house), anti-MLL1 (Cell Signaling, 14689), anti-MLL2 (generated in-house), anti-MLL3 (generated in-house), anti-MLL4 (generated in-house), anti-RBBP5 (Bethyl Laboratories, A300-109A), anti-HSP90 (Santa Cruz Biotechnology, 7947), and anti-tubulin (Developmental Studies Hybridoma Bank, E7).

Cao K., Collings C.K., Marshall S.A., Morgan M.A., Rendleman E.J., Wang L., Sze C.C., Sun T., Bartom E.T, & Shilatifard A. (2017). SET1A/COMPASS and shadow enhancers in the regulation of homeotic gene expression. Genes & Development, 31(8), 787-801.

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Histone Modification Antibody Panel

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The following antibodies are used in this study: anti-H3K4me1 (Cell Signaling Technology [CST], 5326), anti-H3K4me2 (generated in house), anti-H3K4me3 (CST, 9751), anti-H3K27ac (CST, 8173), H3K27me3 (CST, 9733), anti-MLL3 NT (generated in house), anti-MLL3 MT (generated in house), anti-MLL4 NT (generated in house), anti-MLL4 CT (generated in house), anti-MLL2 (generated in house), anti-SET1A (generated in house), anti-Menin (CST, 6891), anti-MLL1C (CST, 14197), anti-RBBP5 (Bethyl Laboratories, A300-109A), anti-NCOA6 (Bethyl Laboratories, A300-410A), anti-UTX (CST, 33510), anti-PTIP (Bethyl Laboratories, A300-370A), ASH2L (CST, 5019), H3 Ser10-p (CST 53348), CDT1 (CST, 8064), Cyclin B1 (CST, 12231), Geminin (CST, 52508), Cyclin E1 (CST, 20808), Cyclin A2 (CST, 91500), p-cdc2 (CST, 4539), PARP (CST, 9542) Caspase3 (CST, 9662), anti-GART (Santa Cruz Biotechnology, sc-166447), anti-PAICS (Proteintech, 12967-1-AP), Hsp90 (Santa Cruz Biotechnology, sc-13119), and anti-β-tubulin (Developmental Studies Hybridoma Bank, E7). Western blot analysis was performed as previously described (53 (link)).

Zhao Z., Cao K., Watanabe J., Philips C.N., Zeidner J.M., Ishi Y., Wang Q., Gold S.R., Junkins K., Bartom E.T., Yue F., Chandel N.S., Hashizume R., Ben-Sahra I, & Shilatifard A. (2023). Therapeutic targeting of metabolic vulnerabilities in cancers with MLL3/4-COMPASS epigenetic regulator mutations. The Journal of Clinical Investigation, 133(13), e169993.

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Spindle Extraction and Immunoblotting

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The samples were electrophoresed on a SDS-PAGE gel, transferred on nitrocellulose membrane (Amersham) and immunoblotted with anti-MLL C (1:250, Bethyl), anti-WDR5 (1:1000,Bethyl), anti-RbBP5 (1:1000, Bethyl), anti-Kif2A (1:1000, Abcam), anti-GFP (1:1000, Invitrogen), anti-GAPDH (1:10000, Sigma), anti-GST(1:10000, Abcam), anti-H3S10P (abcam), and anti-MLL2 (1:250, Bethyl).
Proteins were detected with Licor-Biosciences imaging system as described previously (Tyagi et al., 2007) using IR Dye 800CW conjugated anti-mouse IgG( Licor Biosciences), IR Dye 680 LT conjugated anti-rabbit IgG ( Licor Biosciences).
Spindle Extraction U2OS cells were seeded in 150 x 25mm tissue culture plates. At 60-70% confluency cells were synchronized by thymidine-nocodazole block as described (Harper, 2005) . The cells were then harvested by mitotic shake off method. The spindle extraction was done as described (Sillje ´and Nigg, 2006) . The spindle extract was subjected to SDS-PAGE followed by immunoblotting.

Ali A., Veeranki S.N., Chinchole A, & Tyagi S. (2017). MLL/WDR5 Complex Regulates Kif2A Localization to Ensure Chromosome Congression and Proper Spindle Assembly during Mitosis. Developmental cell, 41(6).

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